In this Phase II project, BlueInGreen, LLC will build a hypersaturated dissolved ozone (HYDOZTM) unit to deploy in a large pilot-scale study at Springdale Wastewater Treatment Facility (SWWTF) in Springdale, Arkansas. Furthermore, BlueInGreen will collaborate with the University of Arkansas to measure the ability of HYDOZ ozonation to remove refractory microbes and stable chemical contaminants from wastewater effluent in both lab-scale and pilot- scale studies. Phase I results demonstrated the increased efficiency and effectiveness of HYDOZ ozone treatment at bench scale compared to conventional ozonation systems. Phase II research will focus on stable contaminants and refractory microbes that are not removed from wastewater using standard chlorine-based disinfection. We will show that the use of the HYDOZ to treat wastewater at centralized wastewater treatment facilities is a viable method economically and technologically to prevent the release of pharmaceutical residuals and resistant microbes to the environment. This project directly addresses the mission of the National Institute of Environmental Health Sciences to reduce the burden of human disease and dysfunction from the environment and will demonstrate both the efficacy and efficiency of HYDOZ ozonation of wastewater effluent. The specific Phase II objectives are: " Objective 1: Compare cost and efficacy of the HYDOZ to standard disinfection/decontamination technology (BlueInGreen). Milestone 1: The capitol and operating cost of HYDOZ systems is superior to chlorine-based disinfection systems, and based on standard effluent quality parameters, the HYDOZ systems are superior to chlorine-based disinfection systems for treatment of wastewater effluent. " Objective 2: Study HYDOZ destruction of a range of chemical contaminants in wastewater effluent (BlueInGreen). Milestone 2: The HYDOZ ozonation systems will cause a minimum 90% reduction in concentration of at least six chemical contaminants from wastewater effluent. " Objective 3: Study HYDOZ destruction of plasmids and refractory microbes in wastewater effluent (University of Arkansas). Milestone 3: The HYDOZ ozonation systems will cause a minimum 90% reduction in concentration of plasmids and refractory microbes from wastewater effluent. PUBLIC HEALTH RELEVANCE: The goal of this project is to use the hypersaturated dissolved ozone (HYDOZTM) system to remove stable chemical/pharmaceutical residuals and refractory microbes from wastewater. Successful reduction of the occurrence of these contaminants in the aqueous environment will reduce long-term impacts on ecosystems and humans, and reduce the spread of antibiotic resistant microbes. The widespread use of this device will significantly contribute to the reduction of antibiotic resistant pathogens in the human ecosystem and help maintain the efficacy of medically important antibiotics.